Purification of a Modified Flagellar Export Apparatus

改良鞭毛出口装置的纯化

• 批准号: 8394244
• 负责人: JONATHAN L MCMURRY
• 金额: $ 2.29万
• 依托单位: KENNESAW STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8394244
• 关键词: Address; Affect; Affinity; Award; Bacterial Infections; Binding; Biochemical; Biological Assay; Biological Models; Biological Process; Biomedical Research; Biosensing Techniques; Chemotaxis; Collaborations; Complement; Complex; Detergents; Development; Dissociation; Dysentery; Environment; Event; Filament; Flagella; Funding; Genes; Genetic; Goals; In Vitro; Institution; Interferometry; Kinetics; Liposomes; Measures; Membrane; Mentors; Methods; Molecular; Molecular Biology; Optics; Pathogenesis; Positioning Attribute; Preparation; Process; Progress Reports; Protein Export Pathway; Proteins; Protons; Publications; Recruitment Activity; Research; Research Personnel; Research Training; Salmonella enterica; Signal Transduction; Structure; Students; Surveys; System; Techniques; Training; Typhoid Fever; United States National Institutes of Health; Variant; Vesicle; Virulence; Work; assay development; career; cell motility; improved; in vivo; instrumentation; kinetosome; meetings; member; nanomachine; pathogenic bacteria; programs; protein protein interaction; reconstitution; retinal rods; stoichiometry; success; tool; undergraduate research

DESCRIPTION (provided by applicant): The bacterial flagellum is a self-assembling, proton-powered rotary nanomachine that drives motility, allowing chemotaxis. It contains a dedicated type III secretion (T3S) apparatus. Homologous T3S systems are used by pathogenic bacteria to effect virulence, underlying many bacterial diseases such as typhoid fever and bacterial dysentery. Using Salmonella enterica as a model system, we will investigate dynamic interactions among flagellar proteins that allow membrane translocation of export substrates through use of molecular biology, genetics and biochemical techniques including optical biosensing. The specific aims are to complete a kinetic survey of interactions among export apparatus proteins using optical biosensing as the primary tool, to examine higher-than-pairwise interactions that better reflect in vivo dynamics and to further the development of an in vitro T3S assay. Eventual development of an in vitro export assay will be of tremendous significance because it would allow us to examine hypotheses addressing T3SS function currently out of reach. Since the flagellar export apparatus is highly similar to virulence T3S systems, the project will contribute significantly to our understanding of bacterial pathogenesis. AREA program goals of enhancing the research environment and exposing students to meritorious research will be well-served by this project.

描述（由申请人提供）：细菌鞭毛是一种自组装的，质子驱动的旋转纳米机械，可驱动运动性，允许趋化性。它包含专用III型分泌（T3S）设备。病原细菌使用同源T3S系统来作用毒力，这是许多细菌性疾病（例如伤寒和细菌痢疾）的基础。我们将使用沙门氏菌作为模型系统，我们将研究鞭毛蛋白之间的动态相互作用，这些蛋白质可以通过使用分子生物学，遗传学和生化技术（包括光学生物敏化）来允许出口底物的膜易位。具体的目的是使用光学生物传感作为主要工具来完成对导出设备蛋白之间的相互作用的动力学调查，以检查更好地反映体内动力学的高度相互作用，并进一步发展体外T3S分析。最终开发体外出口测定法将具有巨大的意义，因为它将允许我们检查解决当前无法触及的T3SS功能的假设。由于鞭毛出口设备与毒力T3S系统高度相似，因此该项目将对我们对细菌发病机理的理解产生重大贡献。该项目将为增强研究环境和使学生暴露于有功研究的领域计划目标。